Humoral and Cellular Immunology in Human Prostate Cancer: Plasma Cells and T and B Lymphocytes / Inmunología Humoral y Celular en el Cáncer de Próstata Humano: Células Plasmáticas y Linfocitos T y B

SUMMARY: In solid and malignant tumors, innate and adaptive immunity are combined in antitumor responses. This study aimed to analyze the activation of plasma cells and the correlation between the infiltration of B and T lymphocytes with the degree of malignancy or Gleason grade in human prostate biopsies diagnosed with cancer. Prostate cancer biopsies were obtained from the Clinical Hospital of Universidad de Chile (n=70), according to the bioethical norms of the institution. Histological sections of 5µm thickness were processed for immunohistochemistry with primary antibodies against BL and total TL (HRP/DAB). Recognition and quantification were performed under a Leica DM750 optical microscope. Microsoft Excel and GraphPad software were used for the statistical study. Correlation coefficient (Pearson) and mean comparison tests (Kruskal-Wallis and Dunn) and p≤ 0.05 were developed. B and T lymphocyte populations were inversely interregulated in prostate cancer (Gleason) (r= -0.46). Their relationship with Gleason grade is variable according to lymphocyte type (LB vs. Gleason r= -0.0.47 and LT vs. Gleason r= -0.21). Histological diagnosis of prostate cancer correlates with a predominance of LT. The malignancy of the pathology correlates with a predominance of LTs, according to the Gleason grade. The increased knowledge of B and T lymphocyte infiltration and plasma cell activation could be used to better target clinical trials on treatments based on immune system responses. Immunotherapy could be a new paradigm to apply better antitumor therapy strategies.

En tumores sólidos y malignos, la inmunidad innata y adaptativa se combinan en respuestas antitumorales. Este estudio tuvo como objetivo analizar la activación de células plasmáticas y la correlación entre la infiltración de linfocitos B y T con el grado de malignidad o grado de Gleason en biopsias de próstata humana diagnosticadas con cáncer. Las biopsias de cáncer de próstata se obtuvieron del Hospital Clínico de la Universidad de Chile (n=70), de acuerdo con las normas bioéticas de la institución. Secciones histológicas de 5 µm de espesor fueron procesadas para inmunohistoquímica con anticuerpos primarios contra LB y LT total (HRP/DAB). El reconocimiento y las cuantificaciones se realizaron bajo un microscopio óptico Leica DM750. Para el estudio estadístico se utilizaron los programas Microsoft Excel y GraphPad. Se desarrollaron pruebas de coeficiente de correlación (Pearson) y comparación de medias (Kruskal-Wallis y Dunn) y p≤ 0.05. Los resultados muestran que las poblaciones de linfocitos B y T están inversamente interreguladas en el cáncer de próstata (r= -0,4578). Su relación con el grado de Gleason es variable según el tipo de linfocito (LB vs Gleason r= -0,47* y LT vs Gleason r= -0,21). Se concluye que la malignidad del cáncer de próstata se correlaciona con un predominio de LT, versus el grado de Gleason. El mayor conocimiento de la infiltración de linfocitos B y T y la activación de células plasmáticas podría aprovecharse para una mejor orientación de ensayos clínicos en tratamientos basados en las respuestas del sistema inmunitario. La inmunoterapia podría ser un nuevo paradigma para aplicar mejores estrategias de terapias antitumorales.

Real-world nailfold videocapillaroscopy in a referral centre in north-western Colombia: A real-life cohort

Introduction: Nailfold videocapillaroscopy is a non-invasive tool for the assessment of peripheral microcirculation, the main indication is the study of Raynaud's phenomenon, poorly standardized outside of this context. There is no clear information in real-life about the rea-sons for referral, the presence of clinical findings of autoimmune diseases, the frequency of patterns of autoantibodies, and specific capillaroscopic findings. Objective: The purpose of this survey is to describe the sociodemographic, clinical, paraclinical, and angioscopy findings of a cohort of subjects referred to a capillaroscopy service in North-western Colombia. Methods: A retrospective study was conducted, from 2015 to 2018. Categorical variables were expressed in frequency and percentage and quantitative variables in mean and standard deviation or median with interquartile range, depending on the distribution of the data. Results: A total of 318 capillaroscopies were performed for the first time. The main referral reason was Raynaud's phenomenon (n = 134; 42.1%). The most frequent baseline capillaroscopic pattern found was normal (n = 123; 38.7%). Of the 12 capillaroscopies that presented a non-specific pattern at a 6-month follow-up, only one (8.3%) progressed to a scleroderma pattern. In the subjects with systemic sclerosis, the most frequent clinical finding was sclerodactyly (n = 34; 37.8%), and 42/44 individuals (95.4%) had positive antinuclear antibodies; the most frequent pattern was centromere (n = 27; 64.3%) Conclusions: In a real-world setting, the main referral reason for capillaroscopy was Raynaud's phenomenon; more than a third of the subjects had normal capillaroscopic findings. Sclerodactyly was the most frequent clinical finding in patients with scleroderma capillaroscopic pattern.

Introducción: La videocapilaroscopia del lecho ungular es una herramienta no invasiva para la evaluación de la microcirculación periférica; la indicación principal es el estudio del fenómeno de Raynaud. Luego de una revisión de la literatura, no hay información clara sobre los motivos de remisión, presencia de hallazgos clínicos de enfermedades autoinmunes, frecuencia de patrones de autoanticuerpos y hallazgos capilaroscópicos específicos. Objetivo: Describir los hallazgos sociodemográficos, clínicos, paraclínicos y capilaroscópicos de sujetos remitidos a un servicio de capilaroscopia en el noroccidente colombiano. Métodos: Estudio retrospectivo de 2015 a 2018. Las variables categóricas se expresaron en frecuencias absolutas y porcentajes, y las variables cuantitativas en media y desviación estándar o mediana con rango intercuartílico, dependiendo de la distribución de los datos. Resultados: Se realizaron 318 capilaroscopias por primera vez. El principal motivo de remisión fue el fenómeno de Raynaud (n = 134; 42,1%). El patrón capilaroscópico basal más frecuente fue el normal (n = 123; 38,7%). De las 12 capilaroscopias que presentaron un patrón no específico en un seguimiento de seis meses, solo una (8,3%) progresó a un patrón de esclerodermia. En los sujetos con esclerosis sistémica, el hallazgo clínico más frecuente fue la esclerodactilia (n = 34; 37,8%), y 42/44 individuos (95,4%) tenían anticuerpos antinucleares positivos; el patrón más frecuente fue el centromérico (n = 27; 64,3%). Conclusiones: La razón principal de remisión para realizar una capilaroscopia fue el fenómeno de Raynaud; más de un tercio de los sujetos tenían hallazgos capilaroscópicos normales. La esclerodactilia fue el hallazgo clínico más frecuente en pacientes con patrón capilaroscópico de esclerodermia.

Analysis of success rate of organoid construction of nasopharyngeal carcinoma by first-day suspension method / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the efficacy of the first-day suspension method for improving the success rate of construction of nasopharyngeal carcinoma-patient derived organoids (NPC-PDO). Methods: The tumor samples of 14 nasopharyngeal carcinoma(NPC) patients, i.e.,13 males and 1 female, with a mean age of 43.0±12.0 years old, were collected from the Affiliated Tumor Hospital of Guangxi Medical University and the First Affiliated Hospital of Guangxi Medical University from January 2022 to July 2022. The tumor samples of 3 patients were digested into single cell suspension and divided into 2 groups, for comparing the efficacy of NPC-PDO construction by the direct inoculation method and the first-day suspension method. The remaining 11 patients were randomized to receive either the direct inoculation method or the first-day suspension method for NPC-PDO construction. The diameter and the number of spheres of NPC-PDO constructed by the two methods were compared by optical microscope; the 3D cell viability detection kit was used to compare the cell viability; the survival rates were compared by trypan blue staining; the success rates of the two construction methods were compared; the number of cases which could be successfully passaged for more than 5 generations and were consistent with the original tissue by pathological examination was counted; and the dynamic changes of cells in suspension overnight were observed by live cell workstation. The independent sample t-test was applied to compare the measurement data of the two groups, and the chi-square test was used to compare the classification data. Results: Compared with the direct inoculation, the diameter and the number of spheres of NPC-PDO constructed by the first-day suspension method were increased, with a higher cell activity, and the success rate of construction was obviously improved (80.0% vs 16.7%, χ2=4.41, P<0.05). In the suspension state, some of the cells aggregated and increased their ability to proliferate. Conclusion: The first-day suspension method can improve the success rate of NPC-PDO construction, especially for those whose original tumor sample size is small.

Ação da luz visível na oxidação da melanina e no descolorimento do cabelo / Action of the visible on the melanin oxidation and hair discoloration

O dano capilar causado pelo descolorimento oxidativo é muito intenso, sendo que dois fatores são responsáveis por essa ação: primeiro, a ação direta e danosa do oxidante em diversas estruturas capilares e segundo, o dano oxidativo primário facilita o dano causado por outros agentes físicos (luz, temperatura) e químicos (tensoativos), que comumente tem ação nos cabelos. Desenvolver conceitos e tecnologias que possam tornar o oxidante específico para a melanina e por conseguinte efetuando o descolorimento sem causar danos ao fio é extremamente desejável. Neste trabalho buscaremos entender de que forma a luz visível pode aumentar a ação do oxidante sem danificar o fio colateralmente. O objetivo principal deste trabalho é demonstrar que é possível utilizar a luz visível, que é absorvida pela melanina, para tornar esse pigmento mais suscetível ao agente oxidante e desta forma, permitir que o descolorimento seja realizado com concentrações pequenas de oxidante. Também almejamos desenvolver métodos de análises por microscopia ótica de fluorescência e de reflexão para mensurar o dano nas estruturas dos fios processados com oxidante e na presença ou ausência da luz

The capillary damage caused by oxidative discoloration is very intense, and two factors are responsible for this action: first, the direct and harmful action of the oxidant on several capillary structures and second, the primary oxidative damage facilitates the damage caused by other physical agents (light, temperature) and chemicals (surfactants), which commonly have action on the hair. Developing concepts and technologies that can make the oxidant specific to melanin and therefore discoloring without causing damage to the hair is extremely desirable. In this work we will try to understand how visible light can increase the oxidant's action without damaging the wire collaterally. The main objective of this work is to demonstrate that it is possible to use visible light, which is absorbed by melanin, to make this pigment more susceptible to the oxidizing agent and, thus, to allow the discoloration to be carried out with small concentrations of oxidizer. We also aim to develop methods of analysis by optical fluorescence and reflection microscopy to measure the damage to the structures of the threads processed with oxidizer and in the presence or absence of light

Satisfaction of medical students in studying histology using virtual and light microscopy: A cross-sectional study

Background@#Medical education has changed as a result of the COVID-19 pandemic. There has been a shift from face to face learning to virtual classes using online learning platforms such as Canvas. These virtual and online alternative methods to medical education brought up concerns about the preparedness of medical students in studying Histology. This study addresses the student’s preference and attitude on the learning of histology using light microscopy vs virtual microscopy. @*Objectives@#The specific objectives of the study are to determine students’ preference, attitudes, and overall satisfaction on the use of light microscopy vs virtual microscopy using a Likert scale. @*Methods@#An enhancement program was conducted by the Department of Anatomy, UP College of Medicine from June 13 to June 17, 2022 among first year medical students. The students were exposed to prosected cadavers, models, specimens, histologic glass slides, and electronic images. During the activity, the second and third floor of Calderon Hall was divided into several stations, each with its own learning outcomes.This is a descriptive cross-sectional study. In all the learning stations, both virtual and light microscopy learning modalities were made available to the students. The student was at liberty to select virtual microscopy, light microscopy or both. In one of the stations, allocated to OS 205 (The study of the anatomy and histology of the thorax), students were randomized to one learning modality (light vs virtual microscopy) and made to identify one predetermined structure. Students answered a short questionnaire that allowed them to express their preference for the modality that was assigned. The questionnaire survey included questions on students’ preference for either light microscopy (LM) or virtual microscopy (VM), ease of use, and satisfaction. A total of five statements were included in the survey questionnaire. All questions in the survey were scored on a 5-point Likert scale (5: strongly disagree, 4: disagree, 3: neutral, 2: agree, and 1: strongly agree). A comments section was also included in the survey to explore students’ experiences of the two learning methods.@*Results@#A total of 160 students participated in the study. Seventy-nine (79) students were randomized to the light microscopy group and 81 one students were randomized to the virtual microscopy group. There were no differences in the demographic characteristics between those randomized to virtual vs light microscopy.There were no differences in the net ratings between those randomised to virtual vs light microscopy in the following domains: 1) ease in looking for structures, 2) ability to identify the structure correctly, 3) method enhancing learning, and 4) overall satisfaction. There was a difference in the net rating between those randomized to virtual vs light microscopy in the domain on quality of the image being easily adjusted (58% vs 97.5%).@*Conclusion@#Medical students who used light microscopes demonstrated a more positive attitude towards its ability to enhance learning and showed greater satisfaction in using this method. Ease in manipulating image quality was better in the light microscopy group. However, the accuracy of identification of histological structures using either platform did not differ. Both virtual and light microscopy are effective learning methods.

Corioamnionitis Histológica Asociada a Muerte Fetal / Histological Chorioamnionitis Associated with Fetal Death

INTRODUCCIÓN. Anualmente ocurren más de 2 millones de muertes fetales a nivel mundial, siendo fundamental el estudio anatomopatológico placentario para disminuir el número de muertes inexplicadas. OBJETIVO. Revisar la literatura existente acerca de corioamnionitis histológica, los criterios para establecer su diagnóstico, su presencia y posible asociación en estudios de causas de muerte fetal. METODOLOGÍA. Se realizaron búsquedas en bases de datos electrónicas para recopilar estudios de causas de muerte fetal que incluyeron corioamnionitis histológica. RESULTADOS. Se encontraron 13 estudios que evaluaron mortalidad fetal y que entre sus causas incluyeron corioamnionitis histológica. DESARROLLO. El estudio microscópico placentario en muertes fetales es esencial al investigar una muerte fetal. Las anomalías placentarias son la causa más común de muerte fetal, la corioamnionitis aguda es la lesión inflamatoria más frecuente. Se detallaron los criterios más relevantes para definir corioamnionitis aguda histológica pero aún no se establece un consenso. Estudios de causas de muerte fetal en años recientes han reportado corioamnionitis histológica entre 6,3% y 41,3% de casos. Las alteraciones inflamatorias del líquido amniótico son una causa importante de muerte fetal, siendo la corioamnionitis la más frecuente en este grupo. CONCLUSIÓN. En estudios para determinar las causas de muerte fetal se evidenció corioamnionitis aguda histológica en hasta el 41,3% de casos, por lo que podría estar asociada a dicho evento. Sin embargo, es necesario establecer un sistema de estadiaje de corioamnionitis histológica mediante un panel de expertos a nivel mundial.

INTRODUCTION. Annually more than 2 million fetal deaths occur worldwide, being fundamental the placental anatomopathological study to reduce the number of unexplained deaths. OBJECTIVE. To review the existing literature on histological chorioamnionitis, the criteria to establish its diagnosis, its presence and possible association in studies of causes of fetal death. METHODOLOGY. Electronic databases were searched to collect studies of causes of fetal death that included histologic chorioamnionitis. RESULTS. Thirteen studies were found that evaluated fetal mortality and that included histologic chorioamnionitis among their causes. DEVELOPMENT: Placental microscopic study in fetal deaths is essential when investigating a fetal death. Placental abnormalities are the most common cause of fetal death, acute chorioamnionitis being the most frequent inflammatory lesion. The most relevant criteria for defining histologic acute chorioamnionitis have been detailed but consensus has not yet been established. Studies of causes of fetal death in recent years have reported histologic chorioamnionitis in between 6,3% and 41,3% of cases. Inflammatory changes in the amniotic fluid are an important cause of fetal death, with chorioamnionitis being the most frequent in this group. CONCLUSIONS. In studies to determine the causes of fetal death, histological acute chorioamnionitis was evidenced in up to 41,3% of cases, so it could be associated with this event. However, it is necessary to establish a histological chorioamnionitis staging system by means of a worldwide panel of experts.

Applying a unified model of fiber dissection, tractography, microscopic anatomy and plastination techniques for basic neuroanatomy education: Hacettepe University experience / Aplicación de un modelo unificado de disección de fibras, tractografía, anatomía microscópica y técnicas de plastinación para la educación básica en neuroanatomía: experiencia de la Universidad Hacettepe

SUMMARY: Anatomy education has gathered together a great many of many new modalities and was modified from classical lecture-based and laboratory practice system to the blended modules. In the scope of the present study, we develop a new, practical, cost- effective and efficient three dimensional (3D) educational model, which aimed to be helpful for the detection and better understanding of basic neuroanatomy education. Tractographic imaging, fiber dissection, microscopic anatomy and plastination techniques were applied to the white matter regions of the two brains. After the photographs that were taken were converted to 3D images, the specimens were plastinated. By way of establishing an educational model as a whole, we applied it to 202 second-year medical students. The students were separated into two groups when they attended to the theoretical lecture. Group 1 took the classical laboratory education; on the other hand, Group 2 received the newly designed educational model. Pre and post-tests were introduced to each group before and after laboratory sessions, respectively. The success scores were put to comparison. The average achievement scores of each group showed increase significantly (p<0.05) after the laboratory sessions, besides the increase in the post-test results of Group 2 was more statistically significant (p<0.05). Consequently, this new educational model enriched by newly designed unified methods could be regarded as useful for grasping and improving the basic neuroanatomy knowledge.

La educación en anatomía ha reunido una gran cantidad de nuevas modalidades, modificándose el sistema clásico de la práctica del laboratorio y de las clases basadas en conferencias, hacia los módulos combinados. En el ámbito del presente estudio, desarrollamos un modelo educativo tridimensional (3D) nuevo, práctico, rentable y eficiente, que pretendía ser útil para la detección y una mejor comprensión de la educación básica en neuroanatomía. Se tomaron imágenes tractográficas, disección de fibras, anatomía microscópica y técnicas de plastinación en los cerebros. Después de convertir las fotografías que se tomaron en imágenes 3D, se plastinaron los especímenes. A modo de establecer un modelo educativo en su conjunto, lo aplicamos a 202 estudiantes de segundo año de medicina. Los estudiantes fueron separados en dos grupos cuando asistieron a la clase teórica. El Grupo 1 tomó la educación clásica de laboratorio; por su parte, el Grupo 2 recibió el nuevo modelo educativo diseñado para el estudio. Se introdujeron pruebas previas y posteriores a cada grupo, antes y después de las sesiones de laboratorio. Se compararon las puntuaciones. Los puntajes promedio de rendimiento de cada grupo mostraron un aumento significativo (p<0,05) después de las sesiones de laboratorio. Además, se obtuvo un aumento en los resultados positivos, posteriores a la prueba del Grupo 2, siendo estadísticamente significativo (p<0,05). En consecuencia, este modelo educativo, enriquecido por métodos unificados de nuevo diseño, podría considerarse útil para captar y mejorar los conocimientos básicos de neuroanatomía.

Aislación y caracterización de las células epiteliales del amnios / Isolation and characterization of amniotic ephitelial cells

RESUMEN: Las células epiteliales del amnios (hAECs) son células madre pluripotenciales; tienen capacidad de diferenciarse en células de las tres capas embrionarias. Como tales, se utilizan en algunas terapias regenerativas en medicina. Este estudio tiene por objetivo describir un protocolo de aislación de las células epiteliales del amnios (hAECs) a partir de placentas humanas de partos por cesárea, así como su caracterización y comportamiento in vitro. Se aislaron hAECs de 20 placentas de partos por cesárea con un protocolo optimizado. Se caracterizaron las células mediante citometría de flujo, microscopia óptica y de fluorescencia, y se evaluó la proliferación de las células mediante MTT a los 1, 3, 5 y 7 días con y sin β-mercaptoetanol en el medio de cultivo. El análisis histológico del amnios mostró un desprendimiento prácticamente completo de las células después de la segunda digestión del amnios. El promedio de células obtenidas fue de 10.97 millones de células por gramo de amnios. Las hAECs mostraron una proliferación limitada, la cual no fue favorecida por la adición de β-mercaptoetanol en el cultivo. Se observó un cambio de morfología espontanea de epitelial a mesenquimal después del cuarto pasaje. Las células epiteliales del amnios pueden ser aisladas con un protocolo simple y efectivo, sin embargo, presentan escasa capacidad proliferativa. Bajo las condiciones de este estudio, la adición de β-mercaptoetanol no favorece la capacidad proliferativa de las células.

SUMMARY: human amnion epithelial cells (hAECs) are pluripotent stem cells; they have the ability to differentiate into cells of the three embryonic layers, and are used in various regenerative therapies in medicine. This study aims to describe a protocol for the isolation of amnion epithelial cells (hAECs) from human placentas from cesarean delivery, as well as their characterization and culture conditions in vitro. hAECs were isolated from 20 cesarean delivery placentas with an optimized protocol. The cells were characterized by flow cytometry, light and fluorescence microscopy, and the proliferation of the cells was evaluated by MTT at 1, 3, 5 and 7 days with and without β-mercaptoethanol in the culture medium. Histological analysis of the amnion showed a practically complete detachment of the cells of the underlying membrane after the second digestion. The average number of cells obtained was 10.97 million cells per amnion. The hAECs perform a limited proliferation rate, which was not favored by the addition of β-mercaptoethanol in the culture. A spontaneous morphology change from epithelial to mesenchymal morphology is exhibited after the fourth passage. The epithelial cells of the amnion can be isolated with a simple and effective protocol, however, they present little proliferative capacity. Under the conditions of this study, the addition of β-mercaptoethanol does not favor the proliferation of the cells.

Histological analysis of an alkaline based tissue clearing technique for fetal musculoskeletal tissue / Análisis histológico de una técnica de limpieza de tejido de base alcalina para tejido musculoesquelético fetal

SUMMARY: Tissue clearing techniques are frequently used in the observation and description of anatomical structures and pathways without altering the three-dimensional layout of the anatomical specimen. Tissue optical clearing promotes preservation of three-dimensional structures, which allows the study of the internal anatomy in its original position and original spatial interaction. Among these techniques, Potassium Hydroxide (KOH) maceration clearing is one of the most widely used. However, the histological changes of tissue after KOH maceration have yet to be fully understood. Our aim is to describe the microscopical differences between macerated and normal tissue. To better understand said changes, two human fetuses with a gestation period of 16 to 28 weeks were cleared and processed for histological analysis. Microtome slides of the fetuses' lower limbs were obtained and stained with Hematoxylin & Eosin, Periodic Acid Schiff (PAS), and Masson's trichrome with the purpose of observing the histological and macromolecule composition changes in cleared tissue. Remarkable differences at a histological level regarding the composition of the cellular structures, since diaphanized tissues showed a predominance of extracellular matrix composed of collagen fibers with the absence of most of the nucleated cellular tissue. Phospholipid's saponification, nucleic acids degradation and a change on proteins structural properties are the main factors inducing clearing. At the same time, molecular stability of collagen in alkaline conditions allows the specimen to maintain its shape after the process.

RESUMEN: Las técnicas de limpieza de tejido se utilizan con frecuencia en la observación y descripción de estructuras y vías anatómicas sin alterar el diseño tridimensional de la muestra anatómica. El aclaramiento óptico de tejidos promueve la preservación de estructuras tridimensionales, lo que permite el estudio de la anatomía interna en su posición original y la interacción espacial original. Entre estas técnicas, el aclarado por maceración con Hidróxido de Potasio (KOH) es una de las más utilizadas. Sin embargo, los cambios histológicos del tejido después de la maceración con KOH aún no se han entendido por completo. Nuestro objetivo es describir las diferencias microscópicas entre el tejido macerado y el normal. Para entender mejor dichos cambios, dos fetos humanos con un período de gestación de 16 a 28 semanas fueron aclarados y procesados para análisis histológicos. Se obtuvieron microtomos de las extremidades inferiores de los fetos y se tiñeron con hematoxilina y eosina, ácido peryódico de Schiff (PAS) y tricrómico de Masson con el fin de observar los cambios histológicos y de composición de macromoléculas en el tejido aclarado. Diferencias notables a nivel histológico en cuanto a la composición de las estructuras celulares, ya que los tejidos diafanizados mostraban un predominio de matriz extracelular compuesta por fibras de colágeno con ausencia de la mayor parte del tejido celular nucleado. La saponificación de los fosfolípidos, la degradación de los ácidos nucleicos y un cambio en las propiedades estructurales de las proteínas son los principales factores que inducen la depuración. Al mismo tiempo, la estabilidad molecular del colágeno en condiciones alcalinas permite que la muestra mantenga su forma después del proceso.

Cromoblastomicosis: primer caso alóctono tratado en Chile / Chromoblastomycosis: first allochthonous case treated in Chile

Resumen La cromoblastomicosis es una infección fúngica de la piel y del tejido subcutáneo, de evolución crónica, causada por hongos dematiáceos que se caracterizan por presentar melanina en su pared celular. La enfermedad se presenta en todo el mundo, principalmente en regiones tropicales y subtropicales. En Chile, solo hay un reporte de caso humano hace más de 30 años. Se presenta el caso de un varón de 46 años, haitiano, residente en Chile, con placas verrucosas en la zona tibial anterior de un año de evolución. El diagnóstico de cromoblastomicosis se confirmó al observar células muriformes en la histopatología y colonias dematiáceas en el cultivo micológico; además, en la miscroscopía directa se observaron conidias compatibles con Fonsecaea spp. Luego de seis meses de tratamiento con antimicóticos sistémicos y crioterapia, se logró la remisión completa de las lesiones.

Abstract Chromoblastomycosis is a fungal infection of the skin and subcutaneous tissue, of chronic evolution, caused by dematiaceous fungi. The disease occurs worldwide, mainly in tropical and subtropical regions, but in regions like Chile there is only one report of a human case more than 30 years ago. We present the case of a 46-year-old Haitian man, resident in Chile, with verrucous plaques in the right anterior tibial area of one year of evolution. The diagnosis of chromoblastomycosis was confirmed when muriform cells and dematiaceous colonies were observed in the histopathological analysis and the direct microscopy, respectively. After six months of treatment with systemic antimycotics and cryotherapy, complete remission of the lesions was achieved.

Proctocolectomía total radical laparoscópica por poliposis familiar adenomatosa colorrectal asociada a cáncer de colon izquierdo / Laparoscopic radical total proctocolectomy by colorrectal adenomatous family polyposis associated with left colon cancer

Resumen Objetivo: Presentar el caso de una paciente mujer adulta, joven, con poliposis múltiple, asociado a cáncer colorrectal, evaluando su manejo quirúrgico oncológico. Materiales y Método: Datos e imágenes recopilados de la historia clínica del Hospital de Alta Complejidad Virgen de la Puerta (HACVP) EsSalud - La Libertad. Resultado: Mujer de 33 años que debuta con dolor abdominal en hemiabdomen izquierdo, más deposiciones con sangre. El hallazgo colonoscópico encuentra múltiples pólipos elevados, planos y sésiles en todo el colon, recto y ano compatible con displasias de alto y bajo grado; a nivel de colon izquierdo se halla lesión exofítica, estenosante cuyo resultado fue adenocarcinoma infiltrante moderadamente diferenciado. Por el gran riesgo de malignidad en todo el intestino grueso, incluyendo canal anal, se le realiza proctocolectomía total laparoscópica más ileostomía terminal. Discusión: La poliposis adenomatosa familiar (PAF) es un síndrome cuyo abordaje quirúrgico va desde una colectomía total con anastomosis ileorrectal, proctocolectomía con ileostomía terminal y proctocolectomía total con Pouch y anastomosis ileoanal. Conclusión: Individualizar el caso, sobre la mejor opción quirúrgica a adoptar para un adecuado manejo oncológico.

Aim: To present the case of a young adult female patient with multiple polyposis associated with colorectal cancer, evaluating her surgical oncological management. Materials and Method: Data and images collected from the clinical history of the Hospital de Alta Complejidad "Virgen de la Puerta" (HACVP) EsSalud - La Libertad. Result: 33-year-old woman presenting with abdominal pain in the left hemiabdomen plus bloody stools. At colonoscopy multiple raised, flat and sessile polyps throughout the colon, rectum, and anus compatible with high-and low-grade dysplasias; an exophytic, stenosing lesion was found in the left colon, the result of which was moderately differentiated infiltrating adenocarcinoma. Due to the great risk of malignancy in the entire large intestine including the anal canal, a total laparoscopic proctocolectomy plus terminal ileostomy was performed. Discussion: Familial adenomatous polyposis (FAP) is a syndrome whose surgical approach ranges from a total colectomy with ileorectal anastomosis, proctocolectomy with terminal ileostomy and total proctocolectomy with Pouch and ileoanal anastomosis. Conclusión: Individualize the case, regarding the best surgical option to adopt for an adequate oncological management.

Ocorrência de perigos físicos em alimentos / Occurrence of physical hazards in food

A ocorrência de matérias estranhas com potencial perigo físico em alimentos é um grande desafio para a indústria alimentícia, e a análise microscópica é provavelmente o instrumento mais útil para detectá-las e identificá-las. Considerando a escassez de dados sobre os perigos físicos em produtos alimentícios no Brasil, o objetivo deste estudo foi descrever essa ocorrência nos produtos analisados no Núcleo de Morfologia e Microscopia do Centro de Alimentos do Laboratório Central do Instituto Adolfo Lutz (IAL), em São Paulo, no período de 2008 a 2020. Os resultados revelaram que, das 7.221 amostras de alimentos analisadas, 89 (1,2%) estavam em desacordo com a legislação em vigor, por conterem perigos físicos. De acordo com a categoria de alimentos, a ocorrência foi maior para bebidas (43%), seguida de cereais, farinha e farelo (22%). Quanto ao tipo de matéria estranha, os plásticos (duros e flexíveis) foram os mais frequentes (48%), seguidos dos metais (15%) entre as partículas perigosas detectadas nas amostras. A detecção e identificação de perigos físicos, por meio de análises microscópicas, contribuem para a segurança e qualidade dos produtos alimentícios oferecidos à população (AU).

The occurrence of physical hazards in food is a great challenge for the food industry, and microscopic analysis is probably the most useful instrument to detect and identify them. Considering the scarcity of data on physical hazards in food products in Brazil, the aim of this study was to describe this occurrence in the products analyzed at the Nucleus of Morphology and Microscopy of the Food Center of the Adolf Lutz Institute's Central Laboratory (IAL) in São Paulo, from 2008 to 2020. Results revealed that of the 7,221 analyzed food samples, 89 (1.2%) did not comply with the legislation due to the presence of physical hazards. According to the food category, the occurrence was higher in beverages (43%), followed by cereals, yeasts, flour and bran (22%). Regarding the type of foreign matter, among the dangerous particles detected, the most frequently found were plastics (48%) (hard and flexible), followed by metals (15%). The detection and identification of physical hazards by microscopic analysis contributes to the safety and quality of food products offered to the public. (AU)

Retinal ganglion cell topography and spatial resolving power of eye in the native chicken of Bangladesh / Topografía de células ganglionares de la retina y poder de resolución espacial del ojo en el pollo nativo de Bangladesh

SUMMARY: The distribution of retinal ganglion cells (RGCs) was observed in the retinal wholemount of native chicken (Gallus gallus domestricus) of Bangladesh by using light microscopy. We considered five different anatomic regions (central, nasal, temporal, dorsal, and ventral) of Nissl stained wholemount, and the RGCs were counted, plotted, and measured accordingly. The average area of the retina was 431.75 mm2 while the total number of ganglion cells was 2124431 on average. Only the central area of the retina was the peak density (10400 cells/mm2) area, signifying the acute visual area, whilst the maximum spatial resolving power was 11 cycles/degree. The overall concentration of RGCs gradually declined towards the periphery but the size of cells generally decreased towards centrally. The size of ganglion cell was not uniform (12 to 180 µm2), specifically the central retina, just above the optic disc was packed with tiny-sized cells. The number, topographic distribution, and size of RGCs in native chicken signified their domesticated or terrestrial characters, uneven visual acuteness, and possibly only the central retina was the area for fine vision as the function of RGCs.

RESUMEN: En este studio se observó la distribución de las células ganglionares en la retina (CGR) de pollo nativo (Gallus gallus domesticus) de Bangladesh mediante el uso de microscopía óptica. Consideramos cinco regiones anatómicas diferentes (central, nasal, temporal, dorsal y ventral). Las muestras de CGR se tiñeron con Nissl, posteriormente, se midieron y contó el número de células totales. El área promedio de la retina fue de 431,75 mm2, mientras que el promedio del número total de células ganglionares fue de 2124431. El área central de la retina fue el área de densidad máxima (10400 células / mm2), señalando el área visual aguda, mientras que el poder de resolución espacial máximo fue de 11 ciclos / grado. La concentración general de CGR disminuyó gradualmente hacia la periferia, sin embargo, el tamaño de las células disminuyó hacia el centro. El tamaño de las CGR no fue uniforme (12 a 180 mm2), específicamente en la retina central, por encima del disco óptico, aumentaron significativamente las células pequeñas. El número, la distribución topográfica y el tamaño de las CGR en pollos nativos determinaron las características domésticas o terrestres, agudeza visual desigual y, posiblemente, la función de las CGR, en la retina central era el área de visión fina.

Cord factor producer Mycobacterium abscessus subsp. bolletii in asymptomatic immunocompetent host sputa samples

Abstract We report a case of Mycobacterium abscessus subsp. bolletii colonization in upper respiratory tract of an immunocompetent patient, who was misdiagnosed as tuberculosis by Acid Fast Bacilli (AFB) and cord factor formation observed directly from the sputa culture in liquid medium. This fact reflected a significant impact on the individual case's life and showed the importance to identify the mycobacteria isolated from clinical sample at species level, and to determine the true implication of nontuberculous mycobacteria (NTM) detected in clinical samples.

In vitro antibacterial activities of silver nanoparticles synthesised using the seed extracts of three varieties of Phoenix dactylifera / Atividades antibacterianas in vitro de nanopartículas de prata sintetizadas usando extratos de sementes de três variedades de Phoenix dactylifera

Green synthesis of silver nanoparticles (AgNPs) is an ecofriendly, cost-effective and promising approach for discovery of novel therapeutics. The aim of the current work was to biogenic synthesize, characterize AgNPs using seed extracts of three economically important varieties of date palm (Iklas, Irziz and Shishi), and assess their anti-pathogenic bacterial activities. AgNPs were synthesised then characterised using electron microscopy and Fourier transform infrared analyses. The bactericidal activities of AgNPs against five different bacterial pathogens, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus and Streptococcus pneumoniae, were determined in vitro. In particular, changes in membrane integrity of virulent bacterial strains in response to AgNPs were investigated. Results of lactate dehydrogenase, alkaline phosphatase activity assays, and measurement of membrane potential revealed that the cytotoxic effects of the AgNPs were mainly centred on the plasma membrane of bacterial cells, leading to loss of its integrity and eventually cell death. In conclusion, green synthesis of AgNPs is an efficient, cost-effective and promising strategy to combat virulent antibiotic-resistant strains.

A síntese verde de nanopartículas de prata (AgNPs) é uma abordagem ecologicamente correta, econômica e promissora para a descoberta de novas terapêuticas. O objetivo do presente trabalho foi sintetizar biogênica, caracterizar AgNPs usando extratos de sementes de três variedades economicamente importantes de tamareira (Iklas, Irziz e Shishi) e avaliar suas atividades bacterianas antipatogênicas. AgNPs foram sintetizados e caracterizados usando microscopia eletrônica e análise de infravermelho por transformada de Fourier. As atividades bactericidas de AgNPs contra cinco diferentes patógenos bacterianos, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Staphylococcus aureus resistente à meticilina e Streptococcus pneumoniae, foram determinadas in vitro. Em particular, foram investigadas alterações na integridade da membrana de cepas bacterianas virulentas em resposta a AgNPs. Os resultados da lactato desidrogenase, dos ensaios da atividade da fosfatase alcalina e da medição do potencial de membrana revelaram que os efeitos citotóxicos dos AgNPs estavam principalmente centrados na membrana plasmática das células bacterianas, levando à perda de sua integridade e, eventualmente, à morte celular. A síntese verde de AgNPs é uma estratégia eficiente, econômica e promissora para combater cepas virulentas resistentes a antibióticos.

Research Progress and Application of Digital Pathology Technology in Bone Marrow Morphology / 中国医疗器械杂志

Advances in digital pathology technology have enabled pathologists and laboratory physicians to perform quick, easy, accurate and reproducible analysis of digital images of tissues and cells with the aid of electronic screens and software tools, rather than relying solely on traditional optical microscopy observations. The conventional clinical cytology testing practice is to be replaced by a digital workflow, which includes both digital imaging and image analysis. This article provides an overview of the basic principles of digital pathology techniques, the advances of development of device in cytology digital pathology, and their clinical applications in bone marrow morphology, and existing problems and prospects of digital pathology application in hematology.

Histomorphological structure of the Bursa cloacalis (Bursa of Fabricius) in young Leiothrix lutea: a comparative study using light microscopy and transmission electron microscopy / Estructura histomorfológica de la Bursa cloacalis (Bursa de Fabricius) en jóvenes Leiothrix lutea: un estudio comparativo utilizando microscopía óptica y microscopía electrónica de transmisión

SUMMARY: This study aimed to investigate the microstructure and ultrastructure of the Bursa cloacalis (Bursa of Fabricius) (BC) in young Leiothrix lutea at various days of age (a few days after hatching) using light microscopy and transmission electron microscopy. The bird BC was sampled at 1, 5, 7, and 9 days of age. Immediately after dissection, the structure and integrity of the BC (not degenerative) were retained and the specific temporal features could be visualized precisely. After hematoxylin-eosin staining and uranyl acetate/lead citrate staining, the microstructure and ultrastructure of the BC, respectively, could be observed clearly. The microscopic observations revealed the following: in addition to change in the size of BC or lymphoid follicles, many cavities were found in the BC; the distribution of the lymphoid follicles in Leiothrix lutea was different from that in other birds; and the segregating line between the bursal cortex and medulla became increasingly clear as the age increased. In conclusion, the structural data obtained in this study provides a better understanding of the specific immunological function of the BC in Leiothrix lutea.

RESUMEN: Este estudio tuvo como objetivo investigar la microestructura y ultraestructura de la Bursa cloacalis (BC) en Leiothrix lutea joven unos días después de la eclosión, utilizando microscopía óptica y microscopía electrónica de transmisión. La BC se muestreó a los 1, 5, 7 y 9 días de edad del Leiothrix lutea. Inmediatamente después de la disección, se observó la estructura y la integridad de la CB (no degenerativa) y se pudo visualizar con precisión las características temporales específicas. Después de la tinción con hematoxilina-eosina y con acetato de uranilo /citrato de plomo, pudimos observar claramente la microestructura y ultraestructura de la BC. Las observaciones microscópicas revelaron el cambio en el tamaño de la CB o de los folículos linfoides y además, se encontraron numerosas cavidades en la CB; la distribución de los folículos linfoides en Leiothrix lutea era diferente a la de otras aves; y la línea de segregación entre la corteza bursal y la médula se hizo cada vez más clara a medida que aumentaba la edad. En conclusión, los datos estructurales obtenidos en este estudio proporcionan una mejor comprensión de la función inmunológica específica de la Bursa cloacalis en Leiothrix lutea.

Microscopia endotelial manual y automatizada: estudio comparativo en adultos sin alteraciones corneales / Manual and automated endothelial microscopy in adults without corneal alterations

Objetivo: Comparar los resultados de los parámetros morfológicos y morfométricos del endotelio corneal a través de la microscopia especular a partir de la utilización del conteo manual y automatizado. Métodos: Se realizó un estudio descriptivo, observacional, de corte transversal de serie de casos en adultos sin alteraciones corneales, en el Servicio de Cirugía Refractiva del Instituto Cubano de Oftalmología "Ramón Pando Ferrer", en el periodo comprendido de enero de 2019 a enero de 2020. Se estudiaron un total de 50 adultos (100 ojos), de los cuales 50 fueron ojos derechos y 50 ojos izquierdos. Se emplearon los porcentajes y los números absolutos para resumir las variables cualitativas. En el caso de las cuantitativas, se utilizó la media con su desviación estándar. Resultados: El 46 por ciento fue del sexo femenino y el 54 por ciento del sexo masculino. Los parámetros morfométricos, como la paquimetría central, fue igual en ambos conteos. A su vez, se obtuvieron mayores valores de densidad endotelial con el conteo automatizado. En los parámetros morfológicos se registró que la hexagonalidad fue superior en el conteo manual y el coeficiente de variabilidad en el conteo automatizado. Las diferencias más marcadas resultaron en el tamaño mínimo, en el máximo y en el promedio de las células endoteliales corneales, con variaciones entre los dos conteos. Conclusiones: Queda demostrado que tanto el conteo manual como el automatizado para el estudio del endotelio corneal pueden ser aplicados en la práctica oftalmológica(AU)

Objective: Compare the results of morphological and morphometric parameters of the corneal endothelium using specular microscopy with manual and automated counting. Methods: A cross-sectional observational descriptive study was conducted of a case series of adults without corneal alterations attending the Refractive Surgery Service at Ramón Pando Ferrer Cuban Institute of Ophthalmology from January 2019 to January 2020. A total 50 adults were studied (100 eyes: 50 right and 50 left). Percentages and absolute numbers were used to summarize qualitative variables, whereas quantitative variables were evaluated with means and standard deviations. Results: Of the patients studied, 46 percent were female and 54 percent were male. Morphometric parameters such as central pachymetry were the same in both countings. Automated counting yielded higher endothelial density values. Morphological parameters showed that hexagonality was greater in manual counting, whereas the coefficient of variability was greater in automated counting. The most obvious differences were observed in minimum size, maximum size and average corneal endothelial cells, with variations between the two countings. Conclusions: It was shown that both manual and automated counting may be used for examination of the corneal endothelium in ophthalmic practice(AU)

Influencia de la interpolación sobre la calidad de imágenes de microscopía celular / Influence of Interpolation on the Quality of Cell Microscopy Images

El presente trabajo tiene el propósito de analizar, para el caso particular de imágenes de microscopía celular de eritrocitos de sangre humana, en qué medida puede influir en la calidad de la imagen la aplicación de métodos de interpolación para mejorar la resolución de esta y con qué métodos se podrían obtener resultados satisfactorios. Se aplicaron para su comparación tres métodos de interpolación a las imágenes a color seleccionadas: splines cúbicos, bicúbico y bilineal y se evaluó la eficiencia computacional de estos. Se utilizaron dos factores de reducción de la resolución (2 y 4) por filas y columnas de la imagen digital. Las medidas utilizadas para valorar la calidad de las imágenes interpoladas fueron la relación señal-ruido y el error medio cuadrático, cuyos valores fueron procesados estadísticamente mediante las pruebas de Friedman y de Wilcoxon, esta última como test post-hoc. Los resultados permiten recomendar el método bicúbico de interpolación como el más favorable para este tipo de imágenes ya que fue el de mejor comportamiento entre los utilizados(AU)

The present work has the purpose of analyzing, for the particular case of cell microscopy images of erythrocytes from human blood, to what extent the application of interpolation methods to improve the image resolution can influence the image quality and with which methods satisfactory results might be obtained. Three interpolation methods were applied for their comparison to the selected color images: cubic splines, bicubic and bilinear and their computational efficiency was also evaluated. Two resolution reduction factors (2 and 4) were used for rows and columns of the digital image. The measures used to assess the quality of the interpolated images were the signal-to-noise ratio and the mean square error, whose values were statistically processed using the Friedman and Wilcoxon tests, the latter as a post-hoc test. The results make it possible to recommend the bicubic interpolation method as the most favorable for this type of images since it was the one with the best performance among those used(AU)

Ultrastructural aspects of the skin in lipoid proteinosis (Urbach-Wiethe disease)

Abstract Lipoid proteinosis is a rare autosomal recessive disease, characterized by hyaline deposits of PAS-positive material in tissues due to mutations in the ECM1 gene. This study evaluated the ultrastructure of the skin of a 6-year-old child affected by this condition. The light microscopy identified PAS-positive hyaline deposits, which were more intense in the papillary dermis. Scanning electron microscopy of the dermis showed a compact papillary dermis and fibrillar deposits in the middle dermis. Transmission electron microscopy clearly showed the deposition of fibrillar material in the dermis, forming clusters adherent to elastic fibers, between the collagen bundles and the collagen fibers, and also filling up the cytoplasm of dermal fibroblasts.